Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1992 Mar 15;89(6):2237–2241. doi: 10.1073/pnas.89.6.2237

Malignant transformation of human fibroblast cell strain MSU-1.1 by (+-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo [a]pyrene.

D Yang 1, C Louden 1, D S Reinhold 1, S K Kohler 1, V M Maher 1, J J McCormick 1
PMCID: PMC48632  PMID: 1549589

Abstract

Treatment of MSU-1.1 cells, a near-diploid, karyotypically stable, infinite life-span human fibroblast strain, with (+-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene induced focus formation. Eight independent foci were isolated and the cell strains developed from them were examined for characteristics of malignant cells. Each grew to a higher density in medium containing 1% serum than did the MSU-1.1 cells. Three of the eight grew rapidly in serum-free medium without added growth factors, formed colonies in agarose with diameters of greater than or equal to 120 microns at a frequency of 5-19%, exhibited loss of genetic material, and, when injected into athymic mice, formed sarcomas that reached 6 mm in diameter within 2-3 wk. One produced high-grade sarcomas (progressively growing, invasive tumors exhibiting high mitotic activity); the other two produced low-grade sarcomas (tumors with a lower degree of mitotic activity) that developed focal areas of high-grade malignant cells if left in the animals for greater than 4 wk. A fourth cell strain formed high-grade sarcomas only after 2.5-3 mo, but the tumor-derived cells analyzed showed the same growth properties as the three malignant cell strains described above, exhibited loss of genetic material, and, when reinjected into athymic mice, produced high-grade sarcomas with a short latency period.

Full text

PDF
2237

Images in this article

2238Image
on p.2238
2240Image
on p.2240

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amstad P., Reddel R. R., Pfeifer A., Malan-Shibley L., Mark G. E., 3rd, Harris C. C. Neoplastic transformation of a human bronchial epithelial cell line by a recombinant retrovirus encoding viral Harvey ras. Mol Carcinog. 1988;1(3):151–160. doi: 10.1002/mc.2940010303. [DOI] [PubMed] [Google Scholar]
  2. Aust A. E., Drinkwater N. R., Debien K., Maher V. M., McCormick J. J. Comparison of the frequency of diphtheria toxin and thioguanine resistance induced by a series of carcinogens to analyze their mutational specificities in diploid human fibroblasts. Mutat Res. 1984 Jan;125(1):95–104. doi: 10.1016/0027-5107(84)90036-8. [DOI] [PubMed] [Google Scholar]
  3. Bettger W. J., Boyce S. T., Walthall B. J., Ham R. G. Rapid clonal growth and serial passage of human diploid fibroblasts in a lipid-enriched synthetic medium supplemented with epidermal growth factor, insulin, and dexamethasone. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5588–5592. doi: 10.1073/pnas.78.9.5588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Christian B. J., Loretz L. J., Oberley T. D., Reznikoff C. A. Characterization of human uroepithelial cells immortalized in vitro by simian virus 40. Cancer Res. 1987 Nov 15;47(22):6066–6073. [PubMed] [Google Scholar]
  6. Connell J. R. The relationship between sister chromatid exchange, chromosome aberration and gene mutation induction by several reactive polycyclic hydrocarbon metabolites in cultured mammalian cells. Int J Cancer. 1979 Oct 15;24(4):485–489. doi: 10.1002/ijc.2910240417. [DOI] [PubMed] [Google Scholar]
  7. Costa J., Wesley R. A., Glatstein E., Rosenberg S. A. The grading of soft tissue sarcomas. Results of a clinicohistopathologic correlation in a series of 163 cases. Cancer. 1984 Feb 1;53(3):530–541. doi: 10.1002/1097-0142(19840201)53:3<530::aid-cncr2820530327>3.0.co;2-d. [DOI] [PubMed] [Google Scholar]
  8. Doll R. The epidemiology of cancer. Cancer. 1980 May 15;45(10):2475–2485. doi: 10.1002/1097-0142(19800515)45:10<2475::aid-cncr2820451004>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  9. Fearon E. R., Vogelstein B. A genetic model for colorectal tumorigenesis. Cell. 1990 Jun 1;61(5):759–767. doi: 10.1016/0092-8674(90)90186-i. [DOI] [PubMed] [Google Scholar]
  10. Fry D. G., Milam L. D., Dillberger J. E., Maher V. M., McCormick J. J. Malignant transformation of an infinite life span human fibroblast cell strain by transfection with v-Ki-ras. Oncogene. 1990 Sep;5(9):1415–1418. [PubMed] [Google Scholar]
  11. Hurlin P. J., Maher V. M., McCormick J. J. Malignant transformation of human fibroblasts caused by expression of a transfected T24 HRAS oncogene. Proc Natl Acad Sci U S A. 1989 Jan;86(1):187–191. doi: 10.1073/pnas.86.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kopchick J. J., Cullen B. R., Stacey D. W. Rapid analysis of small nucleic acid samples by gel electrophoresis. Anal Biochem. 1981 Aug;115(2):419–423. doi: 10.1016/0003-2697(81)90027-0. [DOI] [PubMed] [Google Scholar]
  13. McCormick J. J., Maher V. M. Towards an understanding of the malignant transformation of diploid human fibroblasts. Mutat Res. 1988 Jun;199(2):273–291. doi: 10.1016/0027-5107(88)90209-6. [DOI] [PubMed] [Google Scholar]
  14. Morgan T. L., Yang D. J., Fry D. G., Hurlin P. J., Kohler S. K., Maher V. M., McCormick J. J. Characteristics of an infinite life span diploid human fibroblast cell strain and a near-diploid strain arising from a clone of cells expressing a transfected v-myc oncogene. Exp Cell Res. 1991 Nov;197(1):125–136. doi: 10.1016/0014-4827(91)90489-h. [DOI] [PubMed] [Google Scholar]
  15. Mulligan L. M., Matlashewski G. J., Scrable H. J., Cavenee W. K. Mechanisms of p53 loss in human sarcomas. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5863–5867. doi: 10.1073/pnas.87.15.5863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Namba M., Nishitani K., Fukushima F., Kimoto T., Nose K. Multistep process of neoplastic transformation of normal human fibroblasts by 60Co gamma rays and Harvey sarcoma viruses. Int J Cancer. 1986 Mar 15;37(3):419–423. doi: 10.1002/ijc.2910370314. [DOI] [PubMed] [Google Scholar]
  17. Namba M., Nishitani K., Hyodoh F., Fukushima F., Kimoto T. Neoplastic transformation of human diploid fibroblasts (KMST-6) by treatment with 60Co gamma rays. Int J Cancer. 1985 Feb 15;35(2):275–280. doi: 10.1002/ijc.2910350221. [DOI] [PubMed] [Google Scholar]
  18. Reznikoff C. A., Loretz L. J., Christian B. J., Wu S. Q., Meisner L. F. Neoplastic transformation of SV40-immortalized human urinary tract epithelial cells by in vitro exposure to 3-methylcholanthrene. Carcinogenesis. 1988 Aug;9(8):1427–1436. doi: 10.1093/carcin/9.8.1427. [DOI] [PubMed] [Google Scholar]
  19. Rhim J. S., Fujita J., Arnstein P., Aaronson S. A. Neoplastic conversion of human keratinocytes by adenovirus 12-SV40 virus and chemical carcinogens. Science. 1986 Apr 18;232(4748):385–388. doi: 10.1126/science.2421406. [DOI] [PubMed] [Google Scholar]
  20. Rhim J. S., Jay G., Arnstein P., Price F. M., Sanford K. K., Aaronson S. A. Neoplastic transformation of human epidermal keratinocytes by AD12-SV40 and Kirsten sarcoma viruses. Science. 1985 Mar 8;227(4691):1250–1252. doi: 10.1126/science.2579430. [DOI] [PubMed] [Google Scholar]
  21. Rhim J. S., Kim C. M., Arnstein P., Huebner R. J., Weisburger E. K., Nelson Ress W. A. Transformation of human osteosarcoma cells by a chemical carcinogen. J Natl Cancer Inst. 1975 Dec;55(6):1291–1294. doi: 10.1093/jnci/55.6.1291. [DOI] [PubMed] [Google Scholar]
  22. Rhim J. S., Park D. K., Arnstein P., Huebner R. J., Weisburger E. K., Nelson-Rees W. A. Transformation of human cells in culture by N-methyl-N'-nitro-N-nitrosoguanidine. Nature. 1975 Aug 28;256(5520):751–753. doi: 10.1038/256751a0. [DOI] [PubMed] [Google Scholar]
  23. Ryan P. A., Maher V. M., McCormick J. J. Modification of MCDB 110 medium to support prolonged growth and consistent high cloning efficiency of diploid human fibroblasts. Exp Cell Res. 1987 Oct;172(2):318–328. doi: 10.1016/0014-4827(87)90390-9. [DOI] [PubMed] [Google Scholar]
  24. Sanford K. K., Evans V. J. A quest for the mechanism of "spontaneous" malignant transformation in culture with associated advances in culture technology. J Natl Cancer Inst. 1982 Jun;68(6):895–913. [PubMed] [Google Scholar]
  25. Stampfer M. R., Bartley J. C. Induction of transformation and continuous cell lines from normal human mammary epithelial cells after exposure to benzo[a]pyrene. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2394–2398. doi: 10.1073/pnas.82.8.2394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stevens C. W., Manoharan T. H., Fahl W. E. Characterization of mutagen-activated cellular oncogenes that confer anchorage independence to human fibroblasts and tumorigenicity to NIH 3T3 cells: sequence analysis of an enzymatically amplified mutant HRAS allele. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3875–3879. doi: 10.1073/pnas.85.11.3875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stratton M. R., Moss S., Warren W., Patterson H., Clark J., Fisher C., Fletcher C. D., Ball A., Thomas M., Gusterson B. A. Mutation of the p53 gene in human soft tissue sarcomas: association with abnormalities of the RB1 gene. Oncogene. 1990 Sep;5(9):1297–1301. [PubMed] [Google Scholar]
  28. Wang Y. Y., Maher V. M., Liskay R. M., McCormick J. J. Carcinogens can induce homologous recombination between duplicated chromosomal sequences in mouse L cells. Mol Cell Biol. 1988 Jan;8(1):196–202. doi: 10.1128/mcb.8.1.196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wilson D. M., Yang D. J., Dillberger J. E., Dietrich S. E., Maher V. M., McCormick J. J. Malignant transformation of human fibroblasts by a transfected N-ras oncogene. Cancer Res. 1990 Sep 1;50(17):5587–5593. [PubMed] [Google Scholar]
  30. Wunder J. S., Czitrom A. A., Kandel R., Andrulis I. L. Analysis of alterations in the retinoblastoma gene and tumor grade in bone and soft-tissue sarcomas. J Natl Cancer Inst. 1991 Feb 6;83(3):194–200. doi: 10.1093/jnci/83.3.194. [DOI] [PubMed] [Google Scholar]
  31. Yang J. L., Maher V. M., McCormick J. J. Amplification and direct nucleotide sequencing of cDNA from the lysate of low numbers of diploid human cells. Gene. 1989 Nov 30;83(2):347–354. doi: 10.1016/0378-1119(89)90121-2. [DOI] [PubMed] [Google Scholar]
  32. Yang L. L., Maher V. M., McCormick J. J. Error-free excision of the cytotoxic,mutagenic N2-deoxyguanosine DNA adduct formed in human fibroblasts by (+/-)-7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5933–5937. doi: 10.1073/pnas.77.10.5933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yang L. L., Maher V. M., McCormick J. J. Relationship between excision repair and the cytotoxic and mutagenic effect of the 'anti' 7,8-diol-9,10-epoxide of benzo[a]pyrene in human cells. Mutat Res. 1982 Jun;94(2):435–447. doi: 10.1016/0027-5107(82)90306-2. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES